Engineered cannabinoid biomarker(s), in vitro and in vivo testing, and related applications

Abstract

Methods and applications to label, detect and engineer native or recombinant CB1 and CB2 receptors for cannabinoid chemicals are provided. Also disclosed are uses to detect the dynamics and real time pharmacokinetics of cannabinoid receptors, and purification of cannabinoid receptors in vivo (humans, animals etc.) and in vitro (in the cells).

What is claimed is: 
     
         1 . The cannabinoid receptor biomarkers as provided herein having the amino acid sequences or proteins having the same amino acid sequences where one or more amino acids are added, deleted, or substituted and having receptor activity via binding to cannabinoids. 
     
     
         2 . A cannabinoid receptor biomarker protein encoded by a DNA sequence that hybridizes with the DNA sequence or its complementary sequence and having activity to detect cannabinoid receptor activity. 
     
     
         3 . The recombinant DNA sequence encoding the protein of any one of  claims 1 and 2 , wherein said nucleotide sequence consists of SEQ ID NO: 1 and 2 or full length complements thereof; and (b) a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 1 and 2. 
     
     
         4 . A vector comprising the any of the DNA sequences of  claim 3 . 
     
     
         5 . A transformant carrying the vector of  claim 4 . 
     
     
         6 . A method of producing a fusing protein with synthetic amino acid sequences binding to the protein of any one of  claims 1 and 2 , wherein the method comprises cultivating the transformant of  claim 5  ( FIG.  1   ). 
     
     
         7 . Methods and applications of screening compounds having ability to trace activity of cannabinoid receptors of any one of  claims 1 and 2  ( FIGS.  3  and  5   ), wherein the method comprises:
 (a) contacting the proteins of any one of  claims 1 and 2  with a substrate (including, but not limiting to, cannabinoids) all to be bound by these proteins in the presence of a test compound to detect the cannabinoid receptor activity of the protein of any one of  claims 1 and 2 ; 
 (b) comparing the cannabinoid receptor activity detected in step (a) with that detected in the absence of the test compounds and selecting a compound that lowers or increases the activity of cannabinoid receptors of any one of  claims 1 and 2 ; and 
 (c) purifying the cannabinoid receptor of the protein of any one of  claims 1 and 2 . ( FIG.  5   ). 
 
     
     
         8 . Methods of screening compounds in live animals having either enhancing or inhibitory activity of cannabinoid receptor activity of the protein of any one of  claims 1 and 2  ( FIG.  6   ), wherein the method comprises:
 (a) contacting the proteins of any one of  claims 1 and 2  with a substrate to be bound by these proteins in the presence of a test compound to detect the cannabinoid receptor activity of the protein of any one of  claims 1 and 2 ; and 
 (b) comparing the cannabinoid receptor activity detected in step (a) with that detected in the absence of the test compound; and 
 (c) selecting a compound that lowers or increases the activity of cannabinoid receptors of any one of  claims 1 and 2 . 
 
     
     
         9 . A composition comprising at least two biomarkers of cannabinoid receptors, or chemical compositions selected from the group consisting of  claims 1, 2, 6, 7, and 8 . 
     
     
         10 . Drug testing, analysis, labeling, or categorization utilizing the cannabinoid receptor biomarkers in  any of the above claims  or combination(s) thereof. 
     
     
         11 . Testing and or analyses of the dynamics and/or pharmacokinetics of one or more of the cannabinoid receptors described in  any of the above claims  or combination(s) thereof in order to obtain information about the efficacy of a cannabis, or hemp-based drug related to immune response suppression or other diseases ( FIG.  9   ).